A Rationally Designed Semiconducting Polymer Brush for NIR‐II Imaging‐Guided Light‐Triggered Remote Control of CRISPR/Cas9 Genome Editing

Abstract

The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR‐associated protein 9 (Cas9) genome‐editing system has shown great potential in biomedical applications. Although physical approaches, viruses, and some nonviral vectors have been employed for CRISPR/Cas9 delivery and induce some promising genome‐editing efficacy, precise genome editing remains challenging and has not been reported yet. Herein, second near‐infrared window (NIR‐II) imaging‐guided NIR‐light‐triggered remote control of the CRISPR/Cas9 genome‐editing strategy is reported based on a rationally designed semiconducting polymer brush (SPPF). SPPF can not only be a vector to deliver CRISPR/Cas9 cassettes but also controls the endolysosomal escape and payloads release through photothermal conversion under laser irradiation. Upon laser exposure, the nanocomplex of SPPF and CRISPR/Cas9 cassettes induces effective site‐specific precise genome editing both in vitro and in vivo with minimal toxicity. Besides, NIR‐II imaging based on SPPF can also be applied to monitor the in vivo distribution of the genome‐editing system and guide laser irradiation in real time. Thus, this study offers a typical paradigm for NIR‐II imaging‐guided NIR‐light‐triggered remote control of the CRISPR/Cas9 system for precise genome editing. This strategy may open an avenue for CRISPR/Cas9 genome‐editing‐based precise gene therapy in the near future.

Aptamer/Peptide‐Functionalized Genome‐Editing System for Effective Immune Restoration through Reversal of PD‐L1‐Mediated Cancer Immunosuppression

“…Non‐viral vectors with a better safety profile usually do not possess sufficient transfection efficiency. [ 7–12 ] The reported non‐viral vectors cover diverse materials such as lipid‐like materials, [ 13,14 ] cell exosomes, [ 15 ] polymers and their hybrids, [ 16–22 ] nucleic acid based materials, [ 23 ] and metal containing materials. [ 24–26 ] Nevertheless, in spite of extensive studies on functionalization of non‐viral vectors, the studies on the vectors for delivery of genome editing plasmids are relatively rare.…”

Grafted semiconducting polymer amphiphiles for multimodal optical imaging and combination phototherapy

“…Another study for the regulation of CRISPR/Cas9 gene editing by SPA was conducted by Chen et al 139 They synthesized SPA34 with uorinated polyethylenimine as the side chain to load CRISPR/Cas9 plasmids. A NIR-II uorescent SP was used as the backbone of SPA34 so that the location of SPA34 can be realtime monitored by NIR-II uorescence imaging, and the quantum yield of SPA34 was 0.23%.…”

CRISPR/Cas9 Editing: Sparking Discussion on Safety in Light of the Need for New Therapeutics

“…[107][108][109][110][111][112] An exciting novel approach was recently demonstrated by Li et al by using a semiconducting polymer brush (SPPF) vector for delivery of the CRISPR/Cas9 cassette. 113 This delivery strategy allows for near-infrared window (NIR-II) real-time imaging, and guidance of the complex in vivo and on NIR laser irradiation is capable of inducing localized gene editing at the target destination. Using this technology, the Cas9 can be targeted and visualized in the body, but the laser irradiation acts similar to a remote control to turn editing on when it reaches its destination.…”